Subsequent effect of subacute T-2 toxicosis on spermatozoa, seminal plasma and testosterone production in rabbits

Pannon White (n=12) male rabbits (weight: 4050 to 4500 g, age: 9 months) received 2 ml of a suspension containing purified T-2 toxin by gavage for 3 days. The daily toxin intake was 4 mg/animal (0.78 to 0.99 mg/kg body weight (BW)). Control animals (n=12) received toxin-free suspension for 3 days. Since a feed-refusal effect was observed on the second day after T-2 administration, a group of bucks (n=10) were kept as controls (no toxin treatment) but on a restricted feeding schedule, that is, the same amount of feed was provided to them as was consumed by the exposed animals. On day 51 of the experiment (i.e. 48 days after the 3-day toxin treatment), semen was collected, and pH, concentration, motility and morphology of the spermatozoa, as well as concentration of citric acid, zinc and fructose in the seminal plasma, were measured. After gonadotropin-releasing hormone (GnRH) analogue treatment, the testosterone level was examined. One day of T-2 toxin treatment dramatically decreased voluntary feed intake (by 27% compared to control, P<0.05) and remained lower (P<0.05) during the first 2 weeks after the withdrawal of the toxin. BW of the contaminated rabbits decreased by 88% on days 17 and 29 compared to controls (P<0.05). No effect of toxin treatment was detected on pH and quantity of the semen or concentration of spermatozoa. The ratio of spermatozoa showing progressive forward motility decreased from 65% to 53% in the semen samples of toxin-treated animals compared to controls (P>0.05). The ratio of spermatozoa with abnormal morphology increased (P<0.05) in the ejaculates collected from the toxin-treated animals. T-2 toxin applied in high doses decreased the concentration of citric acid in seminal plasma (P<0.05). No effect of T-2 toxin on the concentrations of the other seminal plasma parameters (fructose and zinc) was observed. T-2 toxin decreased the basic testosterone level by 45% compared to control (P<0.01) and resulted in lower (P<0.05) GnRH-induced testosterone concentration. Feed restriction, that is, less nutrient intake, resulted in more morphologically abnormal spermatozoa in the semen, but it did not cause significant loss in BW, motility of the spermatozoa, composition of the seminal plasma or testosterone concentration--its effect needs further examination.     

Acute Phase Response in Dairy Cows with Experimentally Induced Escherichia coli Mastitis

Six Finnish Ayrshire cows were challenged intramammarily with 1500 CFU of Escherichia coli (E. coli) into single udder quarters, and the challenge was repeated into contralateral quarters 3 weeks later. All cows received flunixine meglumine once, and 3 of them were also treated with enrofloxacin. At the 2nd challenge, treatments were changed vice versa. The development of mastitis was followed by monitoring of systemic and local clinical signs, and with serial milk and serum samples. Intramarnmary challenge with E. coli produced clinical mastitis in all cows, the severity of the disease varying greatly between the animals. No significant changes between the 2 treatment regimens or sequent challenges were found for any of the clinical parameters. The response of each cow followed the same pattern after both challenges; three of the cows became mildly and the other 3 either moderately or severely affected. Two severely affected cows had to be euthanized because of severe mastitis. Serum haptoglobin and amyloid-A concentrations peaked 2–3 days after bacterial challenge. Serum haptoglobin did not correlate with the severity of the disease. Serum amyloid-A rose gradually in the severely affected cows, and significant differences were found between severely versus moderately or mildly affected cows at day 4. Serum tumor necrosis factor alpha concentrations increased only in the severely affected cows. Serum Cortisol response was prolonged in the severely diseased animals, and was significantly lower after the second challenge. Serum nitrite/nitrate concentration increased in the severely affected cows. This indicated excess nitric oxide production during acute E. coli mastitis. Strongly decreased milk production, and high bacterial growth in the infected quarters were best predictors for the outcome from acute E coli mastitis.     

Factors influencing ovarian activity and sexual behavior of postpartum mares under farm conditions

Management of the postpartum period is one of the most important factors of stud farm medicine. In horses, owing to the long gestation period, the time from parturition to repeat conception needs be short to maintain an optimal yearly foaling interval. For this reason the features of postpartum ovarian activity and sexual behavior were studied under farm conditions. During 2 consecutive breeding seasons, 107 mares on 5 commercial horse farms were monitored after parturition by regular teasing, transrectal ultrasonography and blood sampling for progesterone. Foalings took place from January 1 to June 15. Body condition scoring was carried out within 5 d and at 60 to 65 d after parturition. The first ovulation occurred within 20 d after foaling in 84.1% (90/107) of the mares. The mean intervals from foaling to the first and second ovulations were 17.8 +/- 1.6 d (+/- SEM) and 40.9 +/- 2.7 d (+/- SEM), respectively. The mean intervals from parturition to the first and second ovulation (P < 0.001), the interovulatory interval (P < 0.01), the second follicular phase (P < 0.001), and the time until the first overt estrus (P < 0.01) were significantly longer in mares foaling before the vernal equinox. In the beginning of the breeding season the intervals from parturition to the first ovulation (P < 0.01), to the second ovulation (P < 0.01), and to the first overt estrus (P < 0.001) were significantly longer for primiparous mares than for multiparous animals. There was a tendency for an increased interovulatory interval and for a longer second follicular phase in mares with decreased body condition after parturition (P = 0.069, P = 0.089, respectively). Suckling and breed had no effect on postpartum ovarian activity. We concluded that under field conditions the resumption of cyclic ovarian activity and sexual behavior in mares after foaling are strongly affected by the season of parturition and parity. In some cases, body condition change and other factors may also play a role in influencing postpartum reproductive function.     

Factors affecting plasma progesterone concentration and the retrospective determination of time of ovulation in cyclic mares

Factors influencing plasma progesterone concentration were investigated in seven mares. Two-phase logistic curves were fitted (r=0.98) to plasma progesterone concentrations of blood samples collected once daily. In addition to the effect of time (P<0.001), there were differences (P<0.01) among mares in the peak height of the progesterone plateau and in the (area under the curve) AUC. Plasma progesterone concentrations were higher (P<0.001) after a multiple versus single ovulation. There was an effect of season (P<0.001), but no significant effect of luteal morphology. The retrospective determination of time of ovulation was carried out using a linear model on the seven mares and 25 additional mares. Linear regression on the measured values or on the ratio to the average concentration from D5 to D10, was calculated with the day of cycle between D0 and D4. The ovulation date was then calculated using both of these equations, whether blood sampling was performed twice or thrice weekly on 25 postpartum mares. The accuracy to predict day of ovulation (+/- 1 day) ranged from 88 to 97%. In conclusion, the retrospective estimation of time of ovulation in mares was possible, although the technique had some limitations.     

Postpartal ovarian activity of healthy cows and those affected by subclinical metabolic disorders

The postpartal ovarian activity was studied in healthy cows and in those affected by subclinical fatty liver disease and subclinical primary ketosis. Twenty-six dairy cows, 4–7 years old, with not less than 5000 l milk yield in the preceding lactation, were monitored for the following blood parameters: aspartate-amino-transferase (AST)/glutamicoxalacetic transaminase (GOT), albumin, total protein, total lipid, cholesterol, non-esterified fatty acid (NEFA) and triglyceride levels, glucose and ketones. The ratios albumin : total protein and NEFA : triglyceride were also calculated. Blood samples were collected once a week between days 255 and 260 of gestation and days 1–15 postpartum, and subsequently on the 30th, 45th and 60th days postpartum. Based on the biochemical parameters, the cows were assigned in retrospect to three groups: (1) healthy (n = 5), (2) subclinical fatty liver disease (n = 6), and (3) subclinical primary ketosis (n = 10). The postpartal ovarian activity was assessed by rectal palpation and by milk progesterone levels, determined by radioimmunoassay in samples collected 2–3 times weekly between days 5 and 70 postpartum.As early as 10–18 days postpartum, ovarian activity was evident in healthy cows. This was confirmed by the progesterone profiles indicating three regular ovarian cycles with normal hormone levels within 70 days postpartum. All five healthy cows were detected in estrus and only one failed to conceive after insemination.The six cows with subclinical fatty liver disease had no regular cycles and only a few small follicles were palpable on their ovaries. The milk progesterone fluctuated around the basal level (? 1.6 nmol/l). The first Graafian follicles appeared as late as days 30–35 postpartum and the luteal function commenced only between days 38 and 60, as indicated by palpation of corpora lutea and a concomitant rise in milk progesterone. The progesterone peaks were at the lower physiological limits characteristic of luteal function.The ovarian activity of the ten cows with subclinical primary ketosis was intermediate between that of healthy and fatty liver cows. The first follicles and corpora lutea were somewhat delayed, and the progesterone levels fluctuated considerably during the luteal phase. One cow in this group resumed ovarian inactivity during the 70-day postpartal observation period.     

Bacterial complications of postpartum uterine involution in cattle

The bacterial contamination of the postpartum uterus is a frequent finding which by itself does not disturb the anatomical and histological restoration of tubular genital tract. The improper balance between uterine infection and the intrauterine antimicrobial self-defence mechanisms, however, often results in complications, such as puerperal metritis, clinical endometritis, pyometra and subclinical endometritis. After reviewing the bacteriology of uterine involution, and the predisposing factors for its bacterial complications, this paper defines the different clinical forms, and summarizes their pathology, furthermore, the recent progress in diagnostic considerations and principles of current treatments for these diseases of bovine genitals.     

Ovarian consequences of low dose peroral Fusarium (T-2) toxin in a ewe and heifer model

The effect of low dose peroral Fusarium produced T-2 toxin intake upon the ovarian function was evaluated in ewes (n = 30; Trial 1) and heifers (n = 7; Trial 2). Half of the ewes and all of the heifers were fed rich, acidosis-inducing concentrate. The 30 ewes were divided into 6 groups of 5 animals each. They were given 0, 0.3 or 0.9 mg/day (0, 5 or 15 ug/kg) purified T-2 toxin per os for 21 days (3x2 factorial design). Four of the 7 heifers were fed 9 mg/day (25 ug/kg) of the same purified T-2 toxin for 20 days while 3 remained untreated. The estrus cycles in all animals were synchronized prior to the trials and the T-2 exposure was started in the mid-luteal phase. The acidic condition in the rumen was estimated by the determination of urinary net acid-base excretion. The ovarian activity was followed with blood sampling for progesterone on alternate days (Trial 1) or with ultrasonography and sampling for progesterone daily (Trial 2). All of the heifers and concentrate-fed ewes showed a compensated acidosis, during first two thirds of T-2 exposure. In Trial 1, ovarian malfunction manifested as lower P4 peak concentration in the midluteal phase, shortening of the CL lifespan and prolonged follicular phases. These malfunctions were detected in 3 and 3 ewes fed concentrate and 0.3 mg and 0.9 mg T-2 toxin. Lower P4 peak concentration was observed in 1 ewe fed regular diet and 0.9 mg T-2 toxin. None of the control and acidotic groups (0 mg T-2), or ewes fed regular diet with 0.3 mg T-2 showed any ovarian malfunction. In Trial 2, after PGF2, administration the ovulation occured later and the plasma progesterone level remained low (< 3 nmol/l) for a longer period in T-2 treated heifers, than their untreated control mates (5.0+/-0.7 vs 3.7+/-0.5 d, P<0.05 and 8.3+/-0.4 vs 6.3+/-0.9 d, P<0.01, respectively). These results show that the peroral T-2 intake can significantly retard the folliculus maturation and ovulation and perhaps the subsequent luteinisation also in ruminants kept on concentrate-rich diet.     

Short estrous cycles and estrous signs after premature ovulations induced with cloprostenol and gonadotropin-releasing hormone in cyclic dairy cows

The aim of the present study was to confirm earlier findings, obtained with a small number of animals, that gonadotropin-releasing hormone (GnRH) can shorten corpus luteum functional life when it is administered 24 h after cloprostenol (PG) treatments given 7-9 days after estrus. In addition, the effects of two treatments, PG alone or PG + GnRH given before mid-diestrus, on signs of estrus were studied. Sixty cows in farm conditions were used in the experiment. Eight days after natural estrus, they were given an intramuscularly (i.m.) treatment of cloprostenol (0.5 mg). The animals were then divided into two groups. One group (n = 25) received an i.m. treatment of gonadorelin (0.1 mg) 24 h after the PG treatment (PG + GnRH group), while another group (n = 35) served as controls without any further treatment (PG group). Estrous signs were recorded. Progesterone concentrations were measured from samples of whole milk. No short cycles were observed in the PG group, whereas 33% of the cows in the PG + GnRH group exhibited premature luteal regression (P < 0.05). Cloprostenol treatment on Day 8 had no effect on the intensity of the estrous signs. Instead, GnRH treatment 24 h after PG treatment weakened the estrous signs significantly (P < 0.01). It is concluded that GnRH administration 24 h after a PG treatment given 8 days after estrus can cause short estrous cycles in some cows on an individual basis.     

Acute-phase response in dairy cows with acute postpartum metritis

The diagnostic value of 2 plasma acute-phase proteins, haptoglobin and alpha1-acid glycoprotein, and plasma N-acetyl-beta-D-glucosaminidase enzyme activity were studied in 29 newly calved dairy cows. Nineteen had developed acute metritis with putrid vaginal discharge within 2 wk after calving; 10 were clinically healthy controls. Plasma haptoglobin concentration remained low in most cows with acute postpartum metritis. Only the 3 most severely affected cows exhibited a strong haptoglobin response. These were later culled due to poor condition and reduced fertility. This suggests that in acute uterine infection a highly increased haptoglobin concentration indicates poor prognosis for repeat conception. Plasma alpha1-acid glycoprotein concentration increased in acute postpartum metritis, the response pattern being less prominent than that for haptoglobin. The alpha1-acid glycoprotein concentrations did not correlate with severity of disease, and, consequently, the capacity of alpha1-acid glycoprotein in differentiating genital infections was relatively poor. The highest alpha1-acid glycoprotein concentrations were detected in cows with retained placenta and/or dystocia. Plasma N-acetyl-beta-D-glucosaminidase activity levels did not differ between the cows with acute postpartum metritis and healthy control cows.     

Conception rate after fixed time insemination following ovsynch protocol with and without progesterone supplementation in cyclic and non-cyclic Mediterranean Italian buffaloes (Bubalus bubalis)

The aim of this study was to test the effect of progesterone supplementation to Ovsynch protocol in cyclic and non-cyclic Mediterranean Italian buffaloes on conception rate after fixed time artificial insemination. From 169 pluriparous buffaloes, 2 groups were identified and subjected to: (1) Ovsynch protocol (OV; n=83) and (2) Ovsynch protocol with the supplementation of progesterone from days 0 to 7 (OV+PROG.; n=86). All cows were inseminated 16-20 h after the second GnRH administration. Within each group, non-cyclic buffaloes were identified (OV=21 and OV+PROG.=20). Overall conception rate was significantly higher in cyclic compared to non-cyclic buffaloes: 43.7% versus 17.0%, respectively, P=0.001. A significant effect of progesterone supplementation on conception rate was observed in non-cyclic buffaloes (30% versus 4.7%, P=0.04) but not in cyclic buffaloes (51.5% versus 35.7%, P=0.077). Collectively, the presence of a large follicle (>or=10 mm) detected at the beginning of the Ovsynch protocol by ultrasound significantly affected conception rate (44% versus 8%, P=0.01). The findings of the present study suggest that (i) progesterone supplementation to the Ovsynch protocol in buffaloes increases conception rate in non-cyclic animals, (ii) the presence of a large follicle at the beginning of the Ovsynch protocol is a determining factor for a successful synchronization of ovulation and high conception rates and (iii) ultrasound monitoring can improve the overall efficiency by selectively identifying more suitable cycling animals carrying a responsive follicle at the time of first GnRH administration.